Characterizing the role of MSRA in pancreatic tumorigenesis

表征 MSRA 在胰腺肿瘤发生中的作用

基本信息

批准号：
10658248
负责人：
Iok In Christine Chio
金额：
$ 46.62万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-17 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10658248
关键词：
Acute Adenosine Amino Acids Back Biochemical Biological Biological Assay Biology CRISPR/Cas technology Cell Survival Cells Chemicals Collaborations Communication Complementary DNA Complex Cysteine Data Doxycycline Enzymes Genetic Genomic Instability Growth In Vitro Invaded KRAS oncogenesis KRAS2 gene Knock-in Knock-out Maintenance Malignant Neoplasms Malignant neoplasm of pancreas Medical center Metabolic Metabolic Pathway Metabolism Methionine Mitochondria Modeling Molecular Mouse Strains Mus Mutate Nature Neoplasm Metastasis Normal tissue morphology Oncogenes Organoids Outcome Oxidation-Reduction Oxidoreductase Pancreas Pancreatic Ductal Adenocarcinoma Patients Peptides Periodicity Post-Translational Protein Processing Predisposition Presbyterian Church Primary Neoplasm Prognosis Protein Biosynthesis Proteins Proteomics Pyruvate Kinase Reactive Oxygen Species Resistance Respiration Role Signal Transduction Site Specimen Structure Sulfhydryl Compounds Sulfur Testing Therapeutic Therapeutically Targetable Tissue Microarray Tissues Translations Tumor Tissue biochemical tools cell behavior cell injury cell motility chemoproteomics clinically relevant clinically significant design ectoATPase extracellular glucose metabolism in vivo inhibitor macromolecule methionine sulfoxide methionine sulfoxide reductase migration mouse model mutant neoplastic cell novel organoid transplantation oxidation pancreatic ductal adenocarcinoma cell pancreatic neoplasm pancreatic tumorigenesis pharmacologic premalignant protein expression transplant model tumor growth tumorigenesis ubiquitin isopeptidase ubiquitin-protein ligase wound healing

项目摘要

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDA) is an aggressive malignancy that remains largely incurable. The KRAS oncogene is mutated in 95% of PDAs and acts as a potent driver of PDA growth and maintenance. Oncogenic KRAS induces a profound rewiring of metabolic pathways that are essential for PDA growth. Interestingly, this rewiring generates metabolic by-products, including reactive oxygen species (ROS), that can potentially oxidize macromolecules, resulting in genomic instability and toxic cellular damage. However, in addition to these indiscriminate activities, ROS can also initiate redox signaling through oxidative post-translational modifications of proteinaceous cysteines, such that toggling between the reduced and oxidized states of cysteine constitutes a “redox switch” that can regulate the structure and/or function of a protein. Indeed, using our pancreatic organoid model (Cell, 2015), we previously showed that oncogenic KRAS promotes pancreatic tumorigenesis in part by regulating the cyclic reduction/oxidation of cysteines on select proteins involved in the translation machinery (Cell, 2016), thereby promoting protein synthesis (Cell, 2016), glucose metabolism, and tumor growth (Nature Communications, 2019). While cysteine is well-recognized for its susceptibility to oxidation, it is often not appreciated that methionine, the other sulfur-containing amino acid, can be readily oxidized to methionine sulfoxide, and reduced by the oxidoreductase, methionine sulfoxide reductase A (MSRA). Using a tissue microarray of patient-derived pancreatic tumors and adjacent normal tissues, we observed a stage-dependent decrease in MSRA protein expression in tumor tissues. Consistently, acute knockout of murine MSRA using CRISPR/Cas9 in premalignant pancreatic organoids provides a migratory advantage in vitro and in vivo. Thus, we hypothesize that redox signaling through oxidative post-translational modification of methionine residues (Met-oxPTM) represents an unexplored mechanism that supports pancreatic metastasis. Herein, we propose to i) define the mechanism regulating MSRA expression in PDA, ii) characterize the functional and clinical significance of oxidized methionine peptides in PDA metastasis, and iii) define and characterize targetable vulnerabilities created by Met-oxPTM in PDA cells.

项目摘要胰腺导管腺癌（PDA）是一种侵略性恶性肿瘤，在很大程度上无法治愈。克拉斯癌基因在95％的PDA中被突变，并成为PDA生长和维护的潜在驱动力。致癌 KRAS引起了对PDA生长至关重要的代谢途径的深刻重新布线。有趣的是，这重新布线会产生代谢副产品，包括活性氧（ROS），可以潜在地氧化大分子，导致基因组不稳定性和有毒的细胞损伤。但是，除了这些不加选择的活动，ROS还可以通过氧化后翻译后启动氧化还原信号传导蛋白质半胱氨酸，使得半胱氨酸的还原和氧化状态之间切换可以调节蛋白质的结构和/或功能的“氧化还原开关”。确实，使用我们的胰腺器官模型（Cell，2015），我们先前表明，致癌KRAS促进胰腺肿瘤发生部分通过计算与翻译有关的选定蛋白上半胱氨酸的环状还原/氧化机械（细胞，2016年），从而促进蛋白质合成（细胞，2016年），葡萄糖代谢和肿瘤生长（自然通讯，2019年）。虽然半胱氨酸因其对氧化的敏感性而被广泛认可，但通常是不欣赏其他含硫的氨基酸的方法，可以很容易地氧化为方法亚氧化甲氧化还原酶甲二氨酸还原酶A（MSRA）还原。使用组织患者衍生的胰腺肿瘤和相邻正常组织的微阵列，我们观察到阶段依赖性肿瘤组织中MSRA蛋白表达的降低。一致地，使用MSRA的急性敲除胰腺前胰腺癌中的CRISPR/CAS9在体外和体内提供了迁徙优势。那，我们假设氧化后的氧化后翻译后修饰氧化物信号传导残基（Met-Oxptm）代表支持胰腺转移的意外机制。在此，我们建议i）定义调查PDA中MSRA表达的机制，ii）表征 PDA转移中氧化蛋白辣椒的功能和临床意义，以及III）定义和表征由PDA细胞中Met-Oxptm创建的可靶向漏洞。